Localization of selected mutations on the protein molecular model.

<p>A) Domain organization of the IT4var60 PfEMP1 protein with underlined the construct used in this study. B) A molecular model of the NTS-DBL1α of IT4var60 was built based on the crystal structure of the NTS-DBL1α domain of PAvarO strain (PDB: 2yk0) [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118898#pone.0118898.ref021" target="_blank">21</a>]. Three 90 degrees orthogonal views of the molecule in the cartoon representation with mutated amino acids depicted in color. Subdomains 1, 2 and 3 are colored in black, light brown and light blue, respectively. C) Three 90 degrees orthogonal views of the molecule surface in the surface charge potential representation. Arrows indicate positively charged patches Blue: positive charge; white neutral charge; red: negative charge. D) Three 90 degrees orthogonal views of the molecule in the surface representation with mutated amino acids depicted in color. Mut A (Y73A, K263E): orange; Mut B (K118E, G384H): green; Mut C (K202A, K206A): yellow; Mut D (K97A, K171A): purple; Mut E (K325A, K327A): red; Mut F (K97A): black; Mut G (K263E): blue; Mut H (K31A, K34A): cyan.</p

Sub-grouping and sub-functionalization of the RIFIN multi-copy protein family-3

<p><b>Copyright information:</b></p><p>Taken from "Sub-grouping and sub-functionalization of the RIFIN multi-copy protein family"</p><p>http://www.biomedcentral.com/1471-2164/9/19</p><p>BMC Genomics 2008;9():19-19.</p><p>Published online 15 Jan 2008</p><p>PMCID:PMC2257938.</p><p></p>ditional file . Columns with Orange-Blue represent RSS; columns with yellow-green represent CSS; columns with Salmon-green represent both RSS and CSS. (B) Plots of Z-scores and U-values, for CSS (red curve) and RSS (blue curve) respectively, according to alignment position. The predicted consensus secondary structure is plotted with pink and green bars representing helices and loops, respectively. The heights of the bars indicate conserved predictions. Arrows correlate the highest scoring shifted sites with secondary structure predictions

Sub-grouping and sub-functionalization of the RIFIN multi-copy protein family-1

<p><b>Copyright information:</b></p><p>Taken from "Sub-grouping and sub-functionalization of the RIFIN multi-copy protein family"</p><p>http://www.biomedcentral.com/1471-2164/9/19</p><p>BMC Genomics 2008;9():19-19.</p><p>Published online 15 Jan 2008</p><p>PMCID:PMC2257938.</p><p></p>deletions). The B-group is further subdivided into B1, B2 and B3 clusters. Stars indicate sequences that group atypically. Bootstrap support, after 1000 replicates, is only shown for the branches separating the different groups, dots at nodes indicate bootstrap values above or equal to 60%

Binding capacity and affinity of recombinant NTS-DBL1α (It4var60) variants to RBCs and ligands.

<p>Binding capacity and affinity of recombinant NTS-DBL1α (It4var60) variants to RBCs and ligands.</p

A highly conserved segmental duplication in the subtelomeres of chromosomes varies in copy number-2

Ation of copy numbers and localization of (green) in 3D7, FCR3 and 7G8. Distribution of fluorescent signals at the rim of the parasite nuclei (blue) confirms the position of the SD at the chromosomal ends.<p><b>Copyright information:</b></p><p>Taken from "A highly conserved segmental duplication in the subtelomeres of chromosomes varies in copy number"</p><p>http://www.malariajournal.com/content/7/1/46</p><p>Malaria Journal 2008;7():46-46.</p><p>Published online 7 Mar 2008</p><p>PMCID:PMC2279139.</p><p></p

Functional activity of the mutated proteins.

<p>Mutant proteins were tested for their ability to bind O⁺ RBCs and disrupt FCR3S1.2 rosettes. A) Representative dot plot of protein binding to RBC at the highest concentration (100μg/ml) from flow cytometric analysis. B) Proteins were tested in serial dilution from 100μg/ml to 3.06μg/ml for binding to RBCs. Results are presented as mean fluorescence intensity (MFI) fold increase over a negative control. Three independent duplicate experiments were performed. C) FCR3S1.2 trophozoite rosettes were mechanically disrupted and allowed to reform in presence of different concentration of mutant proteins. Proteins were tested at different concentration ranging from 400μg/ml to 5μg/ml. Three independent experiments were performed in duplicate, results shown are average ± SEM.</p

Mutated proteins binding and affinity to Heparin-FITC.

<p>Affinity of the mutated recombinant proteins for the ligand heparin was tested by microscale thermophoresis. Heparin FITC was kept at constant concentration of 100 nM while proteins were tested at ranging concentration varying between 0.5 to 60000 nM. Measurements were performed at 50% LED power and MST 60. Results were plotted using GraphPad Prism and K_D calculated using NanoTemper analysis software (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0118898#pone.0118898.t001" target="_blank">Table 1</a>).</p

Expression of recombinant proteins in <i>E</i>. <i>coli</i>.

<p>A) Size exclusion chromatogram showing monomeric nature of wt and mutated proteins. B) The purity and quality of the mutants was assessed by electrophoresis: 2 μg of wt and mutant proteins were run on 12% SDS-PAGE gel under reducing conditions and stained with Coomassie. C) Far CD spectra of the proteins studied herein, showing nearly identical secondary structures for all the mutants. For color coding and mutation see Table I.</p

A highly conserved segmental duplication in the subtelomeres of chromosomes varies in copy number-3

–28 hours post invasion. Data was logtransformed and plotted at four-hour intervals for each particular parasite.<p><b>Copyright information:</b></p><p>Taken from "A highly conserved segmental duplication in the subtelomeres of chromosomes varies in copy number"</p><p>http://www.malariajournal.com/content/7/1/46</p><p>Malaria Journal 2008;7():46-46.</p><p>Published online 7 Mar 2008</p><p>PMCID:PMC2279139.</p><p></p